生物
胚胎发生
转化(遗传学)
再生(生物学)
细胞生物学
植物
转基因作物
体细胞
转基因
嵌合体(遗传学)
耐旱性
微繁殖
外源DNA
转录因子
生殖系
作者
Zongliang Chen,Jiaqi Zhou,Mary Galli,Sessen Daniel Iohannes,Theresa Clark,Juan M. Debernardi,Jorge Dubcovsky,David Jackson,Andrea Gallavotti
摘要
Summary Transformation is an indispensable tool for plant genetics and functional genomics. Although stable transformation in maize is no longer a major obstacle, there remains a need for accessible and efficient methods for academic laboratories. Here, we present the GGB system, a rapid and efficient approach optimized for immature embryo transformation in B104 and other maize lines. This system combines two distinct morphogenetic regulators, the wheat GRF4‐GIF1 chimera and the maize BABY BOOM (BBM) transcription factor (hence the name “GGB”) with a modified QuickCorn protocol, enabling regeneration of transformed maize plantlets in c . 2 months with an efficiency 7‐fold higher than when compared to either morphogenic factor used in isolation. Expression of both regulators did not significantly affect development, eliminating the need to excise them after regeneration. However, transmission of the transgenic GGB construct through pollen was significantly reduced, potentially aiding transgenic line containment. We show that the GGB system is adaptable for CRISPR‐Cas9 editing and reporter line generation. Furthermore, stable GGB transformants exhibited high leaf regeneration capacity via somatic embryogenesis. RNA‐seq time‐course profiling of GGB leaf cultures identified additional factors that could promote regeneration and led to the discovery of asparagine and trehalose as additional media components that significantly enhanced leaf regeneration.
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